Is the level of acceptable risks the same for individuals who were once competent, such as previously competent adults with Alzheimer disease, individuals who are not now but are expected to become competent, such as healthy children, and individuals who are not now and likely never will be competent, such as individuals born with severe cognitive disabilities?
To construct B. subtilis KEE, pLFKEE was transformed into B. subtilis 168 competent cells as previously described [58] and selected on LB solid medium supplemented with chloramphenicol.
To determine whether transduced cells cultured under mineralizing conditions were competent to undergo mineralization, cells were cultured in a mineralization medium in 3% CO2 (see Materials and methods)as previously described [ 8].
If not otherwise stated, bacterial transformation was performed by heat shock with competent cells prepared by using the Inoue Method for Preparation and Transformation of Competent E. coli as previously described [ 41].
Competent P. pastoris X-33 cells were obtained as recommended by the supplier and electroporation of competent cells was performed as previously described [ 22].
Using standard transformation methods, the anti-AIV/scramble vector was co-transformed with the red fluorescent protein (RFP) prokaryotic expression vector, pE2-Crimson (Clontech, Mountain View, USA), into CEQ221 competent E. coli cells as previously described (Bevis and Glick 2002) and live inoculum stocks were generated and enumerated.
B. subtilis and E. coli competent cells were prepared as previously described and antibiotics were added to the media at regular concentrations [86], [91].
Competent cells were prepared as previously described [ 50].
Studies were approved by the competent institutional review boards as previously reported [ 7, 9, 17].
Competent cells were prepared as previously described (Yu et al., 2000) of an MG1655 derivative containing pSIM6, a plasmid that carries the λ-red system (Datta et al., 2006).
